Recent studies indicate that two RS viral nostructural proteins (NS1 and NS2) are coded for by two adjacent genes. By means of positive hybrid selection of viral mRNAs and subsequent in vitro translation of the selected RNAs a recombinant RS viral plasmid (pRSC6) that encoded two viral nonstructural proteins was identified. This plasmid that contained a RS viral insert of about 1,050 bases hybridized to viral mRNAs containing about 500-600 bases as indicated by Northern blot analysis. The insert was sequenced completely and found to contain two different nonoverlapping reading frames that coded for proteins that contained 139 and 124 amino acids respectively. Upstream of the second reading frame there was a nine nucleotide GGGGCAAAT sequence between positions 545-553. The recombinant pRSC6 was thus shown to be a cDNA clone of a bicistronic transcript. Bicistronic transcript(s) of RS virus may originate because of a failure of the viral polymerase to pause at the intergenic region.